LOCUS       sequence-360577-       15343 bp    DNA     circular SYN 14-APR-2025
DEFINITION  pHR-EF1a-ABE8e-hyperdLbCas12a-NLS-mCherry-P2A-Puro-WPRE.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15343)
  AUTHORS   Kempton HR, Love KS, Guo LY, Qi LS
  TITLE     Scalable biological signal recording in mammalian cells using Cas12a
            base editors.
  JOURNAL   Nat Chem Biol. 2022 May 30. pii: 10.1038/s41589-022-01034-2. doi: 
            10.1038/s41589-022-01034-2.
  PUBMED    35637351
REFERENCE   2  (bases 1 to 15343)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat Chem 
            Biol. 2022 May 30. pii: 10.1038/s41589-022-01034-2. doi: 
            10.1038/s41589-022-01034-2."
COMMENT     Sequence Label: pSLQ11438
FEATURES             Location/Qualifiers
     source          1..15343
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1..17)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             107..340
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     promoter        complement(365..383)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(366..383)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(673..692)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(906..925)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1025..1047
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1085..1103)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1171..1275
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1276..1344,1345..2136)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1494..1513)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2307..2895
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2796..2815
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3049..3066
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3136..3156)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3141..3470
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3321..3456
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          4604..4669
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             4799..4819
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    5244..5378
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5281..5300)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5335..5354
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             5546..6179
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    6226..6351
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6848..7081
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7266..7310
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    7595..7712
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     intron          7932..8854
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     8811..8831
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             8921..8941
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             9536..13216
                     /codon_start=1
                     /product="CRISPR-associated protein from Lachnospiraceae 
                     bacterium ND2006 (Zetsche et al., 2015)"
                     /label=LbCpf1
                     /translation="SKLEKFTNCYSLSKTLRFKAIPVGKTQENIDNKRLLVEDEKRAED
                     YKGVKKLLDRYYLSFINDVLHSIKLKNLNNYISLFRKKTRTEKENKELENLEINLRKEI
                     AKAFKGNEGYKSLFKKDIIETILPEFLDDKDEIALVNSFNGFTTAFTGFFRNRENMFSE
                     EAKSTSIAFRCINENLTRYISNMDIFEKVDAIFDKHEVQEIKEKILNSDYDVEDFFEGE
                     FFNFVLTQEGIRVYNAIIGGFVTESGEKIKGLNEYINLYNQKTKQKLPKFKPLYKQVLS
                     DRESLSFYGRGYTSDEEVLEVFRNTLNKNSEIFSSIKKLEKLFKNFDEYSSAGIFVKNG
                     PAISTISKRIFGEWNVIRDKWNAEYDDIHLKKKAVVTEKYEDDRRKSFKKIGSFSLEQL
                     QEYADADLSVVEKLKEIIIQKVDEIYKVYGSSEKLFDADFVLEKSLKKNDAVVAIMKDL
                     LDSVKSFENYIKAFFGEGKETNRDESFYGDFVLAYDILLKVDHIYDAIRNYVTQKPYSK
                     DKFKLYFQNPQFMGGWDKDKETDYRATILRYGSKYYLAIMDKKYAKCLQKIDKDDVNGN
                     YEKINYKLLPGPNKMLPKVFFSKKWMAYYNPSEDIQKIYKNGTFKKGDMFNLNDCHKLI
                     DFFKDSISRYPKWSNAYDFNFSETEKYKDIAGFYREVEEQGYKVSFESASKKEVDKLVE
                     EGKLYMFQIYNKDFSDKSHGTPNLHTMYFKLLFDENNHGQIRLSGGAELFMRRASLKKE
                     ELVVHPANSPIANKNPDNPKKTTTLSYDVYKDKRFSEDQYELHIPIAINKCPKNIFKIN
                     TEVRVLLKHDDNPYVIGIARGERNLLYIVVVDGKGNIVEQYSLNEIINNFNGIRIKTDY
                     HSLLDKKEKERFEARQNWTSIENIKELKAGYISQVVHKICELVEKYDAVIALEDLNSGF
                     KNSRVKVEKQVYQKFEKMLIDKLNYMVDKKSNPCATGGALKGYQITNKFESFKSMSTQN
                     GFIFYIPAWLTSKIDPSTGFVNLLKTKYTSIADSKKFISSFDRIMYVPEEDLFEFALDY
                     KNFSRTDADYIKKWKLYSYGNRIRIFRNPKKNNVFDWEEVCLTSAYKELFNKYGINYQQ
                     GDIRALLCEQSDKAFYSSFMALMSLMLQMRNSITGRTDVDFLISPVKNSDGIFYDSRNY
                     EAQENAILPKNADANGAYNIARKVLWAIGQFKKAEDEKLDKVKIAISNKEWLEYAQTSV
                     KH"
     CDS             13217..13264
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             13316..14023
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(13461..13479)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     13729..13748
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             14033..14089
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             14093..14692
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(14093..14112)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     14589..14609
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    14752..15340
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(14805..14825)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(15223..15234)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(join(15343,1..16))
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gataccgtcg acctcgaggg aattaattcg agctcggtac ctttaagacc aatgacttac
       61 aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt
      121 cactcccaac gaagacaaga tctgcttttt gcttgtactg ggtctctctg gttagaccag
      181 atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc
      241 ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga
      301 tccctcagac ccttttagtc agtgtggaaa atctctagca gcatctagaa ttaattccgt
      361 gtattctata gtgtcaccta aatcgtatgt gtatgataca taaggttatg tattaattgt
      421 agccgcgttc taacgacaat atgtacaagc ctaattgtgt agcatctggc ttactgaagc
      481 agaccctatc atctctctcg taaactgccg tcagagtcgg tttggttgga cgaaccttct
      541 gagtttctgg taacgccgtc ccgcacccgg aaatggtcag cgaaccaatc agcagggtca
      601 tcgctagcca gatcctctac gccggacgca tcgtggccgg catcaccggc gccacaggtg
      661 cggttgctgg cgcctatatc gccgacatca ccgatgggga agatcgggct cgccacttcg
      721 ggctcatgag cgcttgtttc ggcgtgggta tggtggcagg ccccgtggcc gggggactgt
      781 tgggcgccat ctccttgcat gcaccattcc ttgcggcggc ggtgctcaac ggcctcaacc
      841 tactactggg ctgcttccta atgcaggagt cgcataaggg agagcgtcga atggtgcact
      901 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
      961 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     1021 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga
     1081 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     1141 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     1201 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     1261 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     1321 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     1381 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     1441 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     1501 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     1561 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     1621 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     1681 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     1741 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     1801 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     1861 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     1921 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     1981 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     2041 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     2101 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     2161 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     2221 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     2281 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     2341 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     2401 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     2461 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     2521 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     2581 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     2641 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     2701 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     2761 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     2821 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     2881 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     2941 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     3001 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     3061 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     3121 cattaatgca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     3181 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     3241 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     3301 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     3361 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     3421 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagcttggac
     3481 acaagacagg cttgcgagat atgtttgaga ataccacttt atcccgcgtc agggagaggc
     3541 agtgcgtaaa aagacgcgga ctcatgtgaa atactggttt ttagtgcgcc agatctctat
     3601 aatctcgcgc aacctatttt cccctcgaac actttttaag ccgtagataa acaggctggg
     3661 acacttcaca tgagcgaaaa atacatcgtc acctgggaca tgttgcagat ccatgcacgt
     3721 aaactcgcaa gccgactgat gccttctgaa caatggaaag gcattattgc cgtaagccgt
     3781 ggcggtctgt accgggtgcg ttactggcgc gtgaactggg tattcgtcat gtcgataccg
     3841 tttgtatttc cagctacgat cacgacaacc agcgcgagct taaagtgctg aaacgcgcag
     3901 aaggcgatgg cgaaggcttc atcgttattg atgacctggt ggataccggt ggtactgcgg
     3961 ttgcgattcg tgaaatgtat ccaaaagcgc actttgtcac catcttcgca aaaccggctg
     4021 gtcgtccgct ggttgatgac tatgttgttg atatcccgca agatacctgg attgaacagc
     4081 cgtgggatat gggcgtcgta ttcgtcccgc caatctccgg tcgctaatct tttcaacgcc
     4141 tggcactgcc gggcgttgtt ctttttaact tcaggcgggt tacaatagtt tccagtaagt
     4201 attctggagg ctgcatccat gacacaggca aacctgagcg aaaccctgtt caaaccccgc
     4261 tttaaacatc ctgaaacctc gacgctagtc cgccgcttta atcacggcgc acaaccgcct
     4321 gtgcagtcgg cccttgatgg taaaaccatc cctcactggt atcgcatgat taaccgtctg
     4381 atgtggatct ggcgcggcat tgacccacgc gaaatcctcg acgtccaggc acgtattgtg
     4441 atgagcgatg ccgaacgtac cgacgatgat ttatacgata cggtgattgg ctaccgtggc
     4501 ggcaactgga tttatgagtg ggccccggat ctttgtgaag gaaccttact tctgtggtgt
     4561 gacataattg gacaaactac ctacagagat ttaaagctct aaggtaaata taaaattttt
     4621 aagtgtataa tgtgttaaac tactgattct aattgtttgt gtattttaga ttccaaccta
     4681 tggaactgat gaatgggagc agtggtggaa tgcctttaat gaggaaaacc tgttttgctc
     4741 agaagaaatg ccatctagtg atgatgaggc tactgctgac tctcaacatt ctactcctcc
     4801 aaaaaagaag agaaaggtag aagaccccaa ggactttcct tcagaattgc taagtttttt
     4861 gagtcatgct gtgtttagta atagaactct tgcttgcttt gctatttaca ccacaaagga
     4921 aaaagctgca ctgctataca agaaaattat ggaaaaatat tctgtaacct ttataagtag
     4981 gcataacagt tataatcata acatactgtt ttttcttact ccacacaggc atagagtgtc
     5041 tgctattaat aactatgctc aaaaattgtg tacctttagc tttttaattt gtaaaggggt
     5101 taataaggaa tatttgatgt atagtgcctt gactagagat cataatcagc cataccacat
     5161 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
     5221 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     5281 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     5341 tgtccaaact catcaatgta tcttatcatg tctggatcaa ctggataact caagctaacc
     5401 aaaatcatcc caaacttccc accccatacc ctattaccac tgccaattac ctgtggtttc
     5461 atttactcta aacctgtgat tcctctgaat tattttcatt ttaaagaaat tgtatttgtt
     5521 aaatatgtac tacaaactta gtagttggaa gggctaattc actcccaaag aagacaagat
     5581 atccttgatc tgtggatcta ccacacacaa ggctacttcc ctgattagca gaactacaca
     5641 ccagggccag gggtcagata tccactgacc tttggatggt gctacaagct agtaccagtt
     5701 gagccagata aggtagaaga ggccaataaa ggagagaaca ccagcttgtt acaccctgtg
     5761 agcctgcatg ggatggatga cccggagaga gaagtgttag agtggaggtt tgacagccgc
     5821 ctagcatttc atcacgtggc ccgagagctg catccggagt acttcaagaa ctgctgatat
     5881 cgagcttgct acaagggact ttccgctggg gactttccag ggaggcgtgg cctgggcggg
     5941 actggggagt ggcgagccct cagatcctgc atataagcag ctgctttttg cctgtactgg
     6001 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact
     6061 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg
     6121 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag
     6181 tggcgcccga acagggactt gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg
     6241 actcggcttg ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt gagtacgcca
     6301 aaaattttga ctagcggagg ctagaaggag agagatgggt gcgagagcgt cagtattaag
     6361 cgggggagaa ttagatcgcg atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa
     6421 tataaattaa aacatatagt atgggcaagc agggagctag aacgattcgc agttaatcct
     6481 ggcctgttag aaacatcaga aggctgtaga caaatactgg gacagctaca accatccctt
     6541 cagacaggat cagaagaact tagatcatta tataatacag tagcaaccct ctattgtgtg
     6601 catcaaagga tagagataaa agacaccaag gaagctttag acaagataga ggaagagcaa
     6661 aacaaaagta agaccaccgc acagcaagcg gccggccgct gatcttcaga cctggaggag
     6721 gagatatgag ggacaattgg agaagtgaat tatataaata taaagtagta aaaattgaac
     6781 cattaggagt agcacccacc aaggcaaaga gaagagtggt gcagagagaa aaaagagcag
     6841 tgggaatagg agctttgttc cttgggttct tgggagcagc aggaagcact atgggcgcag
     6901 cgtcaatgac gctgacggta caggccagac aattattgtc tggtatagtg cagcagcaga
     6961 acaatttgct gagggctatt gaggcgcaac agcatctgtt gcaactcaca gtctggggca
     7021 tcaagcagct ccaggcaaga atcctggctg tggaaagata cctaaaggat caacagctcc
     7081 tggggatttg gggttgctct ggaaaactca tttgcaccac tgctgtgcct tggaatgcta
     7141 gttggagtaa taaatctctg gaacagattt ggaatcacac gacctggatg gagtgggaca
     7201 gagaaattaa caattacaca agcttaatac actccttaat tgaagaatcg caaaaccagc
     7261 aagaaaagaa tgaacaagaa ttattggaat tagataaatg ggcaagtttg tggaattggt
     7321 ttaacataac aaattggctg tggtatataa aattattcat aatgatagta ggaggcttgg
     7381 taggtttaag aatagttttt gctgtacttt ctatagtgaa tagagttagg cagggatatt
     7441 caccattatc gtttcagacc cacctcccaa ccccgagggg acccgacagg cccgaaggaa
     7501 tagaagaaga aggtggagag agagacagag acagatccat tcgattagtg aacggatctc
     7561 gacggtatcg ccaaatggca gtattcatcc acaattttaa aagaaaaggg gggattgggg
     7621 ggtacagtgc aggggaaaga atagtagaca taatagcaac agacatacaa actaaagaat
     7681 tacaaaaaca aattacaaaa attcaaaatt ttcgggttta ttacagggac agcagagatc
     7741 cagtttggat cgataagctt gatatcgaat tcaattgaac cggtgcctag agaaggtggc
     7801 gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg cctttttccc gagggtgggg
     7861 gagaaccgta tataagtgca gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg
     7921 ccagaacaca ggtaagtgcc gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg
     7981 gcccttgcgt gccttgaatt acttccacct ggctccagta cgtgattctt gatcccgagc
     8041 tggagccagg ggcgggcctt gcgctttagg agccccttcg cctcgtgctt gagttgaggc
     8101 ctggcctggg cgctggggcc gccgcgtgcg aatctggtgg caccttcgcg cctgtctcgc
     8161 tgctttcgat aagtctctag ccatttaaaa tttttgatga cgtgctgcga cgcttttttt
     8221 ctggcaagat agtcttgtaa atgcgggcca ggatctgcac actggtattt cggtttttgg
     8281 gcccgcggcc ggcgacgggg cccgtgcgtc ccagcgcaca tgttcggcga ggcggggcct
     8341 gcgagcgcgg ccaccgagaa tcggacgggg gtagtctcaa gctggccggc ctgctctggt
     8401 gcctggcctc gcgccgccgt gtatcgcccc gccctgggcg gcaaggctgg cccggtcggc
     8461 accagttgcg tgagcggaaa gatggccgct tcccggccct gctccagggg gctcaaaatg
     8521 gaggacgcgg cgctcgggag agcgggcggg tgagtcaccc acacaaagga aaagggcctt
     8581 tccgtcctca gccgtcgctt catgtgactc cacggagtac cgggcgccgt ccaggcacct
     8641 cgattagttc tggagctttt ggagtacgtc gtctttaggt tggggggagg ggttttatgc
     8701 gatggagttt ccccacactg agtgggtgga gactgaagtt aggccagctt ggcacttgat
     8761 gtaattctcc ttggaatttg gcctttttga gtttggatct tggttcattc tcaagcctca
     8821 gacagtggtt caaagttttt ttcttccatt tcaggtgtcg tgaacaacgc gtacggccgc
     8881 caccatgaaa cggacagccg acggaagcga gttcgagtca ccaaagaaga agcggaaagt
     8941 ctctgaggtg gagttttccc acgagtactg gatgagacat gccctgaccc tggccaagag
     9001 ggcacgggat gagagggagg tgcctgtggg agccgtgctg gtgctgaaca atagagtgat
     9061 cggcgagggc tggaacagag ccatcggcct gcacgaccca acagcccatg ccgaaattat
     9121 ggccctgaga cagggcggcc tggtcatgca gaactacaga ctgattgacg ccaccctgta
     9181 cgtgacattc gagccttgcg tgatgtgcgc cggcgccatg atccactcta ggatcggccg
     9241 cgtggtgttt ggcgtgagga actcaaaaag aggcgccgca ggctccctga tgaacgtgct
     9301 gaactacccc ggcatgaatc accgcgtcga aattaccgag ggaatcctgg cagatgaatg
     9361 tgccgccctg ctgtgcgatt tctatcggat gcctagacag gtgttcaatg ctcagaagaa
     9421 ggcccagagc tccatcaact ctggaggatc tagcggagga tcctctggca gcgagacacc
     9481 aggaacaagc gagtcagcaa caccagagag cagtggcggc agcagcggcg gcagcagcaa
     9541 gctggagaag tttacaaact gctactccct gtctaagacc ctgaggttca aggccatccc
     9601 tgtgggcaag acccaggaga acatcgacaa taagcggctg ctggtggagg acgagaagag
     9661 agccgaggat tataagggcg tgaagaagct gctggatcgc tactatctgt cttttatcaa
     9721 cgacgtgctg cacagcatca agctgaagaa tctgaacaat tacatcagcc tgttccggaa
     9781 gaaaaccaga accgagaagg agaataagga gctggagaac ctggagatca atctgcggaa
     9841 ggagatcgcc aaggccttca agggcaacga gggctacaag tccctgttta agaaggatat
     9901 catcgagaca atcctgccag agttcctgga cgataaggac gagatcgccc tggtgaacag
     9961 cttcaatggc tttaccacag ccttcaccgg cttctttcgg aacagagaga atatgttttc
    10021 cgaggaggcc aagagcacat ccatcgcctt caggtgtatc aacgagaatc tgacccgcta
    10081 catctctaat atggacatct tcgagaaggt ggacgccatc tttgataagc acgaggtgca
    10141 ggagatcaag gagaagatcc tgaacagcga ctatgatgtg gaggatttct ttgagggcga
    10201 gttctttaac tttgtgctga cacaggaggg catccgcgtg tataacgcca tcatcggcgg
    10261 cttcgtgacc gagagcggcg agaagatcaa gggcctgaac gagtacatca acctgtataa
    10321 tcagaaaacc aagcagaagc tgcctaagtt taagccactg tataagcagg tgctgagcga
    10381 tcgggagtct ctgagcttct acggccgggg ctatacatcc gatgaggagg tgctggaggt
    10441 gtttagaaac accctgaaca agaacagcga gatcttcagc tccatcaaga agctggagaa
    10501 gctgttcaag aattttgacg agtactctag cgccggcatc tttgtgaaga acggccccgc
    10561 catcagcaca atctccaagc gtatcttcgg cgagtggaac gtgatccggg acaagtggaa
    10621 tgccgagtat gacgatatcc acctgaagaa gaaggccgtg gtgaccgaga agtacgagga
    10681 cgatcggaga aagtccttca agaagatcgg ctccttttct ctggagcagc tgcaggagta
    10741 cgccgacgcc gatctgtctg tggtggagaa gctgaaggag atcatcatcc agaaggtgga
    10801 tgagatctac aaggtgtatg gctcctctga gaagctgttc gacgccgatt ttgtgctgga
    10861 gaagagcctg aagaagaacg acgccgtggt ggccatcatg aaggacctgc tggattctgt
    10921 gaagagcttc gagaattaca tcaaggcctt ctttggcgag ggcaaggaga caaacaggga
    10981 cgagtccttc tatggcgatt ttgtgctggc ctacgacatc ctgctgaagg tggaccacat
    11041 ctacgatgcc atccgcaatt atgtgaccca gaagccctac tctaaggata agttcaagct
    11101 gtattttcag aaccctcagt tcatgggcgg ctgggacaag gataaggaga cagactatcg
    11161 ggccaccatc ctgagatacg gctccaagta ctatctggcc atcatggata agaagtacgc
    11221 caagtgcctg cagaagatcg acaaggacga tgtgaacggc aattacgaga agatcaacta
    11281 taagctgctg cccggcccta ataagatgct gccaaaggtg ttcttttcta agaagtggat
    11341 ggcctactat aaccccagcg aggacatcca gaagatctac aagaatggca cattcaagaa
    11401 gggcgatatg tttaacctga atgactgtca caagctgatc gacttcttta aggatagcat
    11461 ctcccggtat ccaaagtggt ccaatgccta cgatttcaac ttttctgaga cagagaagta
    11521 taaggacatc gccggctttt acagagaggt ggaggagcag ggctataagg tgagcttcga
    11581 gtctgccagc aagaaggagg tggataagct ggtggaggag ggcaagctgt atatgttcca
    11641 gatctataac aaggactttt ccgataagtc tcacggcaca cccaatctgc acaccatgta
    11701 cttcaagctg ctgtttgacg agaacaatca cggacagatc aggctgagcg gaggagcaga
    11761 gctgttcatg aggcgcgcct ccctgaagaa ggaggagctg gtggtgcacc cagccaactc
    11821 ccctatcgcc aacaagaatc cagataatcc caagaaaacc acaaccctgt cctacgacgt
    11881 gtataaggat aagaggtttt ctgaggacca gtacgagctg cacatcccaa tcgccatcaa
    11941 taagtgcccc aagaacatct tcaagatcaa tacagaggtg cgcgtgctgc tgaagcacga
    12001 cgataacccc tatgtgatcg gcatcgccag gggcgagcgc aatctgctgt atatcgtggt
    12061 ggtggacggc aagggcaaca tcgtggagca gtattccctg aacgagatca tcaacaactt
    12121 caacggcatc aggatcaaga cagattacca ctctctgctg gacaagaagg agaaggagag
    12181 gttcgaggcc cgccagaact ggacctccat cgagaatatc aaggagctga aggccggcta
    12241 tatctctcag gtggtgcaca agatctgcga gctggtggag aagtacgatg ccgtgatcgc
    12301 cctggaggac ctgaactctg gctttaagaa tagccgcgtg aaggtggaga agcaggtgta
    12361 tcagaagttc gagaagatgc tgatcgataa gctgaactac atggtggaca agaagtctaa
    12421 tccttgtgca acaggcggcg ccctgaaggg ctatcagatc accaataagt tcgagagctt
    12481 taagtccatg tctacccaga acggcttcat cttttacatc cctgcctggc tgacatccaa
    12541 gatcgatcca tctaccggct ttgtgaacct gctgaaaacc aagtatacca gcatcgccga
    12601 ttccaagaag ttcatcagct cctttgacag gatcatgtac gtgcccgagg aggatctgtt
    12661 cgagtttgcc ctggactata agaacttctc tcgcacagac gccgattaca tcaagaagtg
    12721 gaagctgtac tcctacggca accggatcag aatcttccgg aatcctaaga agaacaacgt
    12781 gttcgactgg gaggaggtgt gcctgaccag cgcctataag gagctgttca acaagtacgg
    12841 catcaattat cagcagggcg atatcagagc cctgctgtgc gagcagtccg acaaggcctt
    12901 ctactctagc tttatggccc tgatgagcct gatgctgcag atgcggaaca gcatcacagg
    12961 ccgcaccgac gtggattttc tgatcagccc tgtgaagaac tccgacggca tcttctacga
    13021 tagccggaac tatgaggccc aggagaatgc catcctgcca aagaacgccg acgccaatgg
    13081 cgcctataac atcgccagaa aggtgctgtg ggccatcggc cagttcaaga aggccgagga
    13141 cgagaagctg gataaggtga agatcgccat ctctaacaag gagtggctgg agtacgccca
    13201 gaccagcgtg aagcacaaaa ggccggcggc cacgaaaaag gccggccagg caaaaaagaa
    13261 aaagggatcc catatggacg gcattggtag tgggagcaac ggcagcagcg gatccatggt
    13321 gagcaagggc gaggaggata acatggccat catcaaggag ttcatgcgct tcaaggtgca
    13381 catggagggc tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc
    13441 ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc
    13501 ctgggacatc ctgtcccctc agttcatgta cggctccaag gcctacgtga agcaccccgc
    13561 cgacatcccc gactacttga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat
    13621 gaacttcgag gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcga
    13681 gttcatctac aaggtgaagc tgcgcggcac caacttcccc tccgacggcc ccgtaatgca
    13741 gaagaagacc atgggctggg aggcctcctc cgagcggatg taccccgagg acggcgccct
    13801 gaagggcgag atcaagcaga ggctgaagct gaaggacggc ggccactacg acgctgaggt
    13861 caagaccacc tacaaggcca agaagcccgt gcagctgccc ggcgcctaca acgtcaacat
    13921 caagttggac atcacctccc acaacgagga ctacaccatc gtggaacagt acgaacgcgc
    13981 cgagggccgc cactccaccg gcggcatgga cgagctgtac aagggaagcg gagctactaa
    14041 cttcagcctg ctgaagcagg ctggagacgt ggaggagaac cctggaccta ccatgaccga
    14101 gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct
    14161 cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat
    14221 cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa
    14281 ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga
    14341 agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct
    14401 ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg
    14461 gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt
    14521 cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc
    14581 cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga
    14641 ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gagcggccgc
    14701 gactctagag tcgacctgca ggcatgcaag cttgatatca agcttatcga taatcaacct
    14761 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
    14821 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
    14881 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
    14941 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
    15001 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
    15061 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
    15121 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt
    15181 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
    15241 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
    15301 cctcagacga gtcggatctc cctttgggcc gcctccccgc atc
//
